Proteomic Analysis of NF90 (Family) Complexes

NF90（家族）复合物的蛋白质组学分析

• 批准号: 6776438
• 负责人: CINDY J ARRIGO
• 金额: $ 2.8万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2005-01-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6776438
• 关键词: DNA repair; DNA replication; RNA binding protein; cell cycle; cell line; cell nucleus; cytoplasm; double stranded RNA; gene expression; postdoctoral investigator; protein isoforms; protein localization; protein protein interaction; protein purification; protein structure; protein structure function; proteomics; two dimensional gel electrophoresis; virus infection mechanism

DESCRIPTION (provided by applicant): The NF90 family of double-stranded RNA binding proteins is generated by alternative splicing of transcripts from the ILF3 gene. Members of this family have been found in several different subcellular locations and interact with varied protein complexes, suggestive of functions in various cellular processes including gene expression, DNA replication and repair, cell growth, and the anti-viral immune response. Although at least 7 unique protein products have been predicted from the cDNAs, it has been established that four family members predominate in human cell lines and that these isoforms exhibit different levels of activity in transient expression assays. These observations raise the possibility that the isoforms participate in specific complexes that display distinct functions within cells. To address this hypothesis, native NF90 containing complexes will be isolated from recently developed, well-characterized stable cell-lines expressing the four isoforms. The composition of the complexes will be established using proteomic techniques. Once the complex compositions are known for asynchronous human cells (Aim 1), a similar approach will be used to investigate the dynamics of the complexes containing different NF90 members as a function of cell cycle phase and viral infection (Aim 2).

描述（由申请人提供）：双链RNA结合蛋白的NF90家族是通过ILF3基因转录物的选择性剪接产生的。 该家族的成员已在多个不同的亚细胞位置被发现，并与各种蛋白质复合物相互作用，暗示其在各种细胞过程中的功能，包括基因表达、DNA 复制和修复、细胞生长和抗病毒免疫反应。 尽管从 cDNA 中预测出至少 7 种独特的蛋白质产物，但已确定四种家族成员在人类细胞系中占主导地位，并且这些亚型在瞬时表达测定中表现出不同水平的活性。 这些观察结果提出了同工型参与特定复合物的可能性，这些复合物在细胞内显示出不同的功能。 为了解决这一假设，将从最近开发的、表征良好的表达四种亚型的稳定细胞系中分离出含有天然 NF90 的复合物。 将使用蛋白质组学技术确定复合物的组成。 一旦了解了异步人类细胞的复杂组成（目标 1），将使用类似的方法来研究包含不同 NF90 成员的复合物的动力学，作为细胞周期阶段和病毒感染的函数（目标 2）。